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Throwing Knives are perfectly balanced for throwing. An anachronistic and eccentric weapon, the throwing knife did not find wide use in the American west. Despite requiring substantial skill to use effectively, the weapon finds itself in capable hands with Marston.

The throwing knife, like the cannonball and tomahawk, is realistically affected by gravity, and goes in an arc when thrown in long distances. Because of this, it is necessary to raise the height of the throw when taking out an enemy further away.

The throwing knife is not very effective in heavy gunfights or long-range combat, but remains a powerful stealth weapon; if used properly it is capable of killing a human opponent or most animals in one hit. This can be useful for a player who wishes to be undetected or is running low on firearm ammunition.

Throwing knives can be used with Dead-Eye Targeting, but only in single player. Title Update 1.03 disabled the use of throwing knives with Dead Eye in Multiplayer. Only one knife can be thrown per Dead Eye sequence. When used this way, throwing knives have far greater range, accuracy and diminished flight-time, massively increasing their usefulness; this can prove quite useful in the mission The Great Mexican Train Robbery. The knife can be used to even bring down birds as far away as dots in the distance.

If used in the Dead-Eye Targeting system and exit before the knife hits the enemy then the knife sometimes disappears and does not hit the target. This results in no kill and also losing the knife. If a thrown knife kills an NPC, the looted body usually returns the knife back to the inventory.

Attacking an enemy with a throwing knife while in melee range will result in a silent, instant kill without using up any of the supply of knives. In addition, a glitch causes the looted body to yield a knife as if it was thrown, actually gaining the player a throwing knife in the process.

Although looting bodies killed with the knife gives the knife back, skinning an animal killed with the knife does not give the knife back.

The throwing knife finds an unexpected niche when it comes to dispatching of enemy horses, as the throwing knives are an instant kill to most animals. This is particularly useful in multiplayer mode, when stunning a pursuer can buy enough time to get the upper hand in battle.

Throwing knives are the only weapon not provided in gang matches.

You can perform a special execution using the knives if you hit an enemy in both legs putting them on their knees. Proceed to attack with melee combat and you will stab the enemy in the neck.

A random glitch occurs when using "Dead Eye" on a hanging rope. The knife will float and spin for a few seconds then make contact with the rope.

The stealth benefit of Throwing knives is useless while playing bounty hunting missions, because as soon as the player kills someone, Marston immediately yells at the bounty to give up. The stealth of the Throwing knife is also somewhat useless in towns, such as Blackwater, because the townspeople can witness the players murders even through the walls/floors of buildings.

In Multiplayer, this is the only Thrown Weapon that cannot be made golden. However, it does appear to have a noticeably golden handle.

Looting an NPC killed with a Throwing Knife will result in the Throwing Knife being added to the player's inventory again. The knife however, will stay in the NPC's body. It happens with the Tomahawk as well.

Hitting certain areas on an NPC with the knife will cause them to stagger a few inches then begin to unleash gunfire on Marston.

When performing an execution on a person with a throwing knife, the person will be stabbed either in front or in their back, depending on which angle the player performed the execution. However, if he performs the 'loot body' function afterwards, then they will receive a throwing knife, which helps getting a throwing knife for free.